3. REAGENT PREPARATION - LAB

Question 1

Where can you find instructions for preparing reagents and quantities used?

Answer 1

Written laboratory protocol

Question 2

What should be documented and matched to test performance in amplification methods?

Answer 2

Lot numbers and working stocks of probes and primers

Question 3

Why should probe sequences used for a given test be recorded?

Answer 3

Because probes for linkage analysis and array technology are periodically updated

Question 4

What is the purpose of an RNA primer in vivo?

Answer 4

Initiates DNA replication

Question 5

What is the purpose of RNA primers in vitro (synthetically made primers)?

Answer 5

Used in DNA amplification

Question 6

What is a critical component of PCR procedures?

Answer 6

Primers

Question 7

What is the lyophilized form of a primer?

Answer 7

freeze dried or powdered form

Question 8

What information is included in the primer product?

Answer 8

Quality, Method of purification, Molecular weight, Number of micrograms of dried primers

Question 9

What is the typical length of oligonucleotide primers in base pairs?

Answer 9

18-25 or 18-30

Question 10

What do probes bind to directly?

Answer 10

Target sequence

Question 11

What is the purpose of probes?

Answer 11

Detection of target sequence or hybridization techniques

Question 12

In what forms are probes supplied?

Answer 12

Solution or dried form

Question 13

How should new working stocks of probes be treated?

Answer 13

As new reagent lots

Question 14

What needs to be done to dried form probes or primers?

Answer 14

Dissolve in solvent to make stock solution

Question 15

What should be done with stock solution to make a working solution?

Answer 15

Dilute it

Question 16

What are analyte-specific reagents (ASRs)?

Answer 16

Probes, primers, antibodies

Question 17

What does analyte-specific reagents detect?

Answer 17

Presence or absence of a particular substance

Question 18

Where are analyte-specific reagents used?

Answer 18

Tissue typing, infectious disease, PCR

Question 19

What are in vitro diagnostic (IVD) devices?

Answer 19

Instruments or systems for diagnosis

Question 20

What are IVD reagent sets used for?

Answer 20

Collecting, preparing, or examining specimens

Question 21

How are IVD devices classified?

Answer 21

CLASS I, CLASS II, CLASS III

Question 22

Reagent categories

Answer 22

ASRs
IVD
IVAT
RUO IUO

Question 23

This category is intended to accommodate the use of promising high complexity technologies facing long clearance processes

Answer 23

In vitro analytical test (IVAT) reagents

Question 24

are not intended for diagnostic use

Answer 24

Research Use Only (RUO)

Question 25

can be used in patient samples with proper institutional review and informed consent

Answer 25

Investigational Use Only (IUO)

Question 26

reagents are used to gather data that may result in the advancement of a product's status

Answer 26

Research Use Only (RUO) and Investigational Use Only (IUO) reagents

Question 27

What is the first step in making reagents and buffers?

Answer 27

Go through the protocol for the experiment and write down all the solutions used.

Question 28

What should you decide about the solution concentration?

Answer 28

Decide whether to make diluted or concentrated solutions.

Question 29

What should you check regarding chemical availability?

Answer 29

Find out if the chemicals you need are available in the lab.

Question 30

What should you check for safety information?

Answer 30

MSDS (Material Safety Data Sheet)

Question 31

What label should you look for on hazardous materials?

Answer 31

Hazardous material classification label

Question 32

Where should you read for warnings about the reagent?

Answer 32

Reagent bottle/container

Question 33

What should you do if there are no warnings on the label?

Answer 33

Look up the material in the Merck Index

Question 34

What if the label is in another language or you can't find a listing?

Answer 34

Consult the manufacturer

Question 35

What is a concern during the analytical phase of work?

Answer 35

Exposure to various hazardous reagents

Question 36

What neurotoxin is used for protein and sequencing gels?

Answer 36

Acrylamide

Question 37

What mutagen intercalates in DNA and is used to label nucleic acids?

Answer 37

Ethidium bromide (EtBr)

Question 38

What highly corrosive substance can burn skin and should be used in a fume hood?

Answer 38

Phenol

Question 39

What reagent, used to inhibit proteinases, can be fatal if swallowed or absorbed through the skin?

Answer 39

Phenylmethyl-sulfonate fluoride (PMSF)

Question 40

What detergent is a burning powder and extremely irritating to nasal passages?

Answer 40

Sodium dodecyl sulfate (SDS)

Question 41

What method is used for sterilizing most buffers?

Answer 41

Autoclave

Question 42

What material are tips and microcentrifuge tubes made of for autoclaving?

Answer 42

High-density polyethylene

Question 43

What should be avoided when autoclaving tips and microcentrifuge tubes?

Answer 43

Proteins

Question 44

What should you check for in solutions before autoclaving?

Answer 44

Acidity or alkalinity

Question 45

What substances should not be autoclaved?

Answer 45

Corrosives, solvents, volatiles, liquids with bleach/formalin/glutaraldehyde, buffers with detergents (e.g., 10% SDS), heat-labile ingredients, DTT, and β-mercaptoethanol

Question 46

Storage of buffers in room temperature

Answer 46

Detergents Ethanol Buffers Concentrated solutions Acid and bases

Question 47

Storage of buffers and solutions in 4C

Answer 47

Bacterial cultures Buffers DNA Media PCR products Serum

Question 48

Storage of buffer and solutions in -20c

Answer 48

Enzymes RNA

Question 49

Storage of buffer and solutions in -70c

Answer 49

Frozen bacterial cultures Lipids